monoclonal antibody analysisAfter the last post on the topic of HPLC analyis of monoclonal antibodies (MAbs) using a newly released hydrophobic interaction chromatography (HIC) column (Thermo ScientificMAbPac HIC-10 column), I am pleased to present another recently released application note featuring a pH-gradient-based method that provides excellent resolution for MAb charge variants.

Glycosylated proteins—including erythropoietins, MAbs, and various hormones—constitute a large portion of major approved therapeutic biological drugs. Sialic acids are important to many biological processes and also have significant effects on the properties of therapeutic proteins. Thus, monitoring protein glycosylation, including sialylation, is important for both glycoprotein characterization and quality control purposes.

Application Note 1092, Separation of Intact Monoclonal Antibody Sialylation Isoforms by pH Gradient Ion-Exchange Chromatography, (downloadable PDF), demonstrates that the pH-gradient approach is more convenient and straightforward than the isoelectric focusing method routinely used for protein sialylation profiling. This new method uses a cation-exchange protein column (Thermo Scientific ProPac SCX-10 LC column) on our biocompatible analytical liquid chromatography system (Thermo Scientific Dionex UltiMate 3000 Dual BioRS LC System). Note that a separate project is underway that focuses on fast separation using a strong cation-exchange column (Thermo Scientific MAbPac SEC-1 Size Exclusion column) that incorporates a smaller particle size.

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Webinar: Improving Monoclonal Antibody Characterization

This on-demand, 1-hour webinar, titled, Achieving hgher throughput and productivity in monoclonal antibody characterization, (link to registration page; webinar start playing once you register for it) could also help you in your work on MAb analysis. One of our experts discusses how to develop robust high throughput methods for MAb characterization.

In case you are working on intact protein analysis, do check out this brochure: Thermo Scientific Solutions for Intact Protein Analysis(downloadable PDF).

Additional Resources

  • Our Chromatography Solution Online Center features many useful and complimentary chromatography tools which can help speed up your analysis. The site is updated on a monthly basis; therefore, do check out the Archives section to see what was previously featured.

If you have questions on the method, the column, or the technique, do enter them in the Comments box below; our experts look forward to hearing from you.